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Lymphoblastoid cell line with B1 cell characteristics established from a chronic lymphocytic leukemia clone by in vitro EBV infection.

Rosén A, Bergh AC, Gogok P, Evaldsson C, Myhrinder AL, Hellqvist E, Rasul A, Björkholm M, Jansson M, Mansouri L, Liu A, Teh BT, Rosenquist R, Klein E - Oncoimmunology (2012)

Bottom Line: Infected cells do not express the growth-promoting set of EBV-encoded genes and therefore they do not yield LCLs, in most experiments.These genes may contribute to establish precursors of CLL clones by regulation of cellular phenotype in the hematopoietic compartment.Expression of CD5/CD20/CD27/CD43 and spontaneous production of natural antibodies may identify the CLL cell as a self-renewing B1 lymphocyte.

View Article: PubMed Central - PubMed

Affiliation: Department of Clinical and Experimental Medicine; Division of Cell Biology; Linköping University; Linköping, Sweden.

ABSTRACT
Chronic lymphocytic leukemia (CLL) cells express the receptor for Epstein-Barr virus (EBV) and can be infected in vitro. Infected cells do not express the growth-promoting set of EBV-encoded genes and therefore they do not yield LCLs, in most experiments. With exceptional clones, lines were obtained however. We describe a new line, HG3, established by in vitro EBV-infection from an IGHV1-2 unmutated CLL patient clone. All cells expressed EBNA-2 and LMP-1, the EBV-encoded genes pivotal for transformation. The karyotype, FISH cytogenetics and SNP-array profile of the line and the patient's ex vivo clone showed biallelic 13q14 deletions with genomic loss of DLEU7, miR15a/miR16-1, the two micro-RNAs that are deleted in 50% of CLL cases. Further features of CLL cells were: expression of CD5/CD20/CD27/CD43 and release of IgM natural antibodies reacting with oxLDL-like epitopes on apoptotic cells (cf. stereotyped subset-1). Comparison with two LCLs established from normal B cells showed 32 genes expressed at higher levels (> 2-fold). Among these were LHX2 and LILRA. These genes may play a role in the development of the disease. LHX2 expression was shown in self-renewing multipotent hematopoietic stem cells, and LILRA4 codes for a receptor for bone marrow stromal cell antigen-2 that contributes to B cell development. Twenty-four genes were expressed at lower levels, among these PARD3 that is essential for asymmetric cell division. These genes may contribute to establish precursors of CLL clones by regulation of cellular phenotype in the hematopoietic compartment. Expression of CD5/CD20/CD27/CD43 and spontaneous production of natural antibodies may identify the CLL cell as a self-renewing B1 lymphocyte.

No MeSH data available.


Related in: MedlinePlus

Figure 1. (A) Human B1 phenotypic markers expressed on HG3. Indirect immunofluorescence analysis with appropriate mAbs followed by FITC-of Cy-ChromeTM-conjugated anti-mouse Ig. Ten thousands events were counted in a FACScan flow cytometer. (B) Competition chemiluminescent ELISA for oxLDL. MDA-LDL competition ELISA was performed for HG3 CLL IgM mAb isolated from cell supernatant and purified by affinity chromatography. Plates were coated with MDA-LDL, then IgM was allowed to react with increasing amounts of soluble competitor MDA-LDL or native LDL (nLDL). B/B0 indicates the ratio between bound (cpm) and bound (cpm) without competitor.
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Figure 1: Figure 1. (A) Human B1 phenotypic markers expressed on HG3. Indirect immunofluorescence analysis with appropriate mAbs followed by FITC-of Cy-ChromeTM-conjugated anti-mouse Ig. Ten thousands events were counted in a FACScan flow cytometer. (B) Competition chemiluminescent ELISA for oxLDL. MDA-LDL competition ELISA was performed for HG3 CLL IgM mAb isolated from cell supernatant and purified by affinity chromatography. Plates were coated with MDA-LDL, then IgM was allowed to react with increasing amounts of soluble competitor MDA-LDL or native LDL (nLDL). B/B0 indicates the ratio between bound (cpm) and bound (cpm) without competitor.

Mentions: All established cell lines strongly expressed CD5. Figure 1A shows the expression of a CD5, CD20, CD27, CD43 markers on the HG3 cells. The cell line showed a weak expression of the monocyte/macrophage marker CD14, and expression of the activation markers CD38 and CD70 (Fig. 1A). The line was also positive for CD19, CD21, CD31, CD69 (not shown). Upon prolonged cultivation, CD5-expression was slightly decreased, whereas the CD14, CD19, CD20, CD21, CD23, CD27, CD31, CD38, CD43, CD69, CD70, IgM and λ−light chain expression were retained.


Lymphoblastoid cell line with B1 cell characteristics established from a chronic lymphocytic leukemia clone by in vitro EBV infection.

Rosén A, Bergh AC, Gogok P, Evaldsson C, Myhrinder AL, Hellqvist E, Rasul A, Björkholm M, Jansson M, Mansouri L, Liu A, Teh BT, Rosenquist R, Klein E - Oncoimmunology (2012)

Figure 1. (A) Human B1 phenotypic markers expressed on HG3. Indirect immunofluorescence analysis with appropriate mAbs followed by FITC-of Cy-ChromeTM-conjugated anti-mouse Ig. Ten thousands events were counted in a FACScan flow cytometer. (B) Competition chemiluminescent ELISA for oxLDL. MDA-LDL competition ELISA was performed for HG3 CLL IgM mAb isolated from cell supernatant and purified by affinity chromatography. Plates were coated with MDA-LDL, then IgM was allowed to react with increasing amounts of soluble competitor MDA-LDL or native LDL (nLDL). B/B0 indicates the ratio between bound (cpm) and bound (cpm) without competitor.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3376971&req=5

Figure 1: Figure 1. (A) Human B1 phenotypic markers expressed on HG3. Indirect immunofluorescence analysis with appropriate mAbs followed by FITC-of Cy-ChromeTM-conjugated anti-mouse Ig. Ten thousands events were counted in a FACScan flow cytometer. (B) Competition chemiluminescent ELISA for oxLDL. MDA-LDL competition ELISA was performed for HG3 CLL IgM mAb isolated from cell supernatant and purified by affinity chromatography. Plates were coated with MDA-LDL, then IgM was allowed to react with increasing amounts of soluble competitor MDA-LDL or native LDL (nLDL). B/B0 indicates the ratio between bound (cpm) and bound (cpm) without competitor.
Mentions: All established cell lines strongly expressed CD5. Figure 1A shows the expression of a CD5, CD20, CD27, CD43 markers on the HG3 cells. The cell line showed a weak expression of the monocyte/macrophage marker CD14, and expression of the activation markers CD38 and CD70 (Fig. 1A). The line was also positive for CD19, CD21, CD31, CD69 (not shown). Upon prolonged cultivation, CD5-expression was slightly decreased, whereas the CD14, CD19, CD20, CD21, CD23, CD27, CD31, CD38, CD43, CD69, CD70, IgM and λ−light chain expression were retained.

Bottom Line: Infected cells do not express the growth-promoting set of EBV-encoded genes and therefore they do not yield LCLs, in most experiments.These genes may contribute to establish precursors of CLL clones by regulation of cellular phenotype in the hematopoietic compartment.Expression of CD5/CD20/CD27/CD43 and spontaneous production of natural antibodies may identify the CLL cell as a self-renewing B1 lymphocyte.

View Article: PubMed Central - PubMed

Affiliation: Department of Clinical and Experimental Medicine; Division of Cell Biology; Linköping University; Linköping, Sweden.

ABSTRACT
Chronic lymphocytic leukemia (CLL) cells express the receptor for Epstein-Barr virus (EBV) and can be infected in vitro. Infected cells do not express the growth-promoting set of EBV-encoded genes and therefore they do not yield LCLs, in most experiments. With exceptional clones, lines were obtained however. We describe a new line, HG3, established by in vitro EBV-infection from an IGHV1-2 unmutated CLL patient clone. All cells expressed EBNA-2 and LMP-1, the EBV-encoded genes pivotal for transformation. The karyotype, FISH cytogenetics and SNP-array profile of the line and the patient's ex vivo clone showed biallelic 13q14 deletions with genomic loss of DLEU7, miR15a/miR16-1, the two micro-RNAs that are deleted in 50% of CLL cases. Further features of CLL cells were: expression of CD5/CD20/CD27/CD43 and release of IgM natural antibodies reacting with oxLDL-like epitopes on apoptotic cells (cf. stereotyped subset-1). Comparison with two LCLs established from normal B cells showed 32 genes expressed at higher levels (> 2-fold). Among these were LHX2 and LILRA. These genes may play a role in the development of the disease. LHX2 expression was shown in self-renewing multipotent hematopoietic stem cells, and LILRA4 codes for a receptor for bone marrow stromal cell antigen-2 that contributes to B cell development. Twenty-four genes were expressed at lower levels, among these PARD3 that is essential for asymmetric cell division. These genes may contribute to establish precursors of CLL clones by regulation of cellular phenotype in the hematopoietic compartment. Expression of CD5/CD20/CD27/CD43 and spontaneous production of natural antibodies may identify the CLL cell as a self-renewing B1 lymphocyte.

No MeSH data available.


Related in: MedlinePlus